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. 2025 Aug 21:14:30-38.
doi: 10.33393/jcb.2025.3564. eCollection 2025 Jan-Dec.

Diagnostic value of carcinoembryonic antigen, cancer antigen 15-3, and cell-free DNA as blood biomarkers in early detection of canine mammary tumor

Diksha Singh  1 , Prashant P Rokade  1 , Neeraj K Gangwar  1 , Mukul G Gabhane  1 , Sunil Malik  1 , Kavisha Gangwar  1 , Shyama N Prabhu  1 , Renu Singh  1 , D D Singh  1 , Sonam Kumari  2 , Soumen Chaudhary  2 , Jitendra K Choudhary  3
Affiliations

Diagnostic value of carcinoembryonic antigen, cancer antigen 15-3, and cell-free DNA as blood biomarkers in early detection of canine mammary tumor

Diksha Singh et al. J Circ Biomark. .

Abstract

Introduction: Blood biomarkers play a crucial role in the diagnosis and prognosis of tumor. The present research was designed to study the diagnostic effect of serum biomarkers, namely carcino-embryonic antigen (CEA), cancer antigen 15-3 (CA15-3), and plasma biomarker viz., circulating cell-free DNA (cfDNA); and their correlation with cytological and histopathological results.

Methods: A total of 60 blood samples were collected. Out of which 36 samples were from the dogs affected with canine mammary tumors, and 24 samples were from the apparently healthy dogs. CEA and CA15-3 were estimated using Sandwich ELISA, and cfDNA was estimated by the ccfDNA kit. A significant Positive correlation was observed between tumor blood biomarker levels, cytology and histopathological grades of the tumors.

Results: We found that CA15-3 can be a more effective serum tumour biomarker than CEA for diagnosing canine mammary gland tumours. This finding showed a positive correlation with the clinical grade of the disease. The concentration of serum markers and cfDNA in animals affected with malignant mammary gland tumours was higher compared to the benign entity of tumours and healthy control groups. The ROC curve analysis revealed that the sensitivity (Se) and specificity (Sp) of CEA and CA15-3 biomarkers improved when used together. IN comparison to healthy controls, canines with both benign and malignant neoplasia showed significantly higher (p < 0.05) cfDNA concentrations.

Conclusion: This study highlights the role of blood tumor biomarkers for routine screening of animals in early diagnosis of tumors, further treatment, and prognosis.

Keywords: CA15-3; CEA; Cytology; Histopathology; Mammary gland tumors; cfDNA.

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Conflict of interest statement

Conflict of interest: The authors declare that they have no potential conflicts of interest.

Figures

FIGURE 1 -
FIGURE 1 -
Cytological observation of different mammary gland tumours in dogs (Geimsa Stain, 1000X). (a) Mildly pleomorphic cells arranged in clusters, Grade I; (b) singly arranged cells with vacuolated cytoplasm, Grade II; (c) Mixed population of pleomorphic cells showing karyokinesis stage, Grade III.
FIGURE 2 -
FIGURE 2 -
Histopathological observation of different mammary gland tumours in dogs (HE Staining, 200X). (a) Carcinoma Mixed Type; (b) Fibroadenoma; (c) comedocarcinoma; (d)tubulo-papillary carcinoma; (e) squamous cell carcinoma; (f) Healthy mammary gland.
FIGURE 3 -
FIGURE 3 -
Expression levels of CEA, CA15-3 in serum and cfDNA in plasma of canine mammary tumor. (a) Serum CEA levels of the malignant tumor group, the benign tumor group, and the healthy control group. (b) Serum CA15-3 levels of the three groups. (c) Plasma cfDNA levels of the three groups. Note: *p < 0.05 showed a significant difference, **p < 0.01 showed an extremely significant difference.
FIGURE 4 -
FIGURE 4 -
The ROC curve of single and combined detection in the diagnosis of canine mammary gland tumor. (a) The ROC curves for the single detection of CEA. (b) The ROC curves for the single detection of CA15-3. (c) The ROC curves of the combined detection of CA15-3+CEA.
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